CN113480677B - Cortex moutan linear alpha-D-1, 4-glucan and preparation method and application thereof - Google Patents
Cortex moutan linear alpha-D-1, 4-glucan and preparation method and application thereof Download PDFInfo
- Publication number
- CN113480677B CN113480677B CN202110883581.8A CN202110883581A CN113480677B CN 113480677 B CN113480677 B CN 113480677B CN 202110883581 A CN202110883581 A CN 202110883581A CN 113480677 B CN113480677 B CN 113480677B
- Authority
- CN
- China
- Prior art keywords
- cortex moutan
- polysaccharide
- glucan
- drying
- crude
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229920001503 Glucan Polymers 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 150000004676 glycans Chemical class 0.000 claims abstract description 62
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 62
- 239000005017 polysaccharide Substances 0.000 claims abstract description 62
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 50
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 claims abstract description 22
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 18
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 18
- 238000001035 drying Methods 0.000 claims abstract description 17
- 238000004108 freeze drying Methods 0.000 claims abstract description 12
- 230000001376 precipitating effect Effects 0.000 claims abstract description 9
- 239000003960 organic solvent Substances 0.000 claims abstract description 7
- 230000004792 oxidative damage Effects 0.000 claims abstract description 5
- 238000001914 filtration Methods 0.000 claims abstract description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 42
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 28
- 238000003756 stirring Methods 0.000 claims description 19
- 239000000843 powder Substances 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 15
- 238000000502 dialysis Methods 0.000 claims description 8
- 238000001556 precipitation Methods 0.000 claims description 8
- 239000003957 anion exchange resin Substances 0.000 claims description 7
- 239000003472 antidiabetic agent Substances 0.000 claims description 7
- 206010012601 diabetes mellitus Diseases 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 5
- 235000013376 functional food Nutrition 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000001641 gel filtration chromatography Methods 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 238000000926 separation method Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 28
- 230000000694 effects Effects 0.000 abstract description 14
- 238000005238 degreasing Methods 0.000 abstract description 8
- 238000010438 heat treatment Methods 0.000 abstract description 7
- 231100000331 toxic Toxicity 0.000 abstract description 4
- 230000002588 toxic effect Effects 0.000 abstract description 4
- 230000001681 protective effect Effects 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 2
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 2
- 238000010992 reflux Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 27
- 239000000047 product Substances 0.000 description 26
- 238000004458 analytical method Methods 0.000 description 22
- 240000005001 Paeonia suffruticosa Species 0.000 description 20
- 235000003889 Paeonia suffruticosa Nutrition 0.000 description 20
- 235000019441 ethanol Nutrition 0.000 description 16
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 14
- 229910052799 carbon Inorganic materials 0.000 description 14
- 229910052739 hydrogen Inorganic materials 0.000 description 13
- 239000001257 hydrogen Substances 0.000 description 13
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 12
- 238000012512 characterization method Methods 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 10
- 238000001514 detection method Methods 0.000 description 10
- 102100023374 Forkhead box protein M1 Human genes 0.000 description 9
- 101000907578 Homo sapiens Forkhead box protein M1 Proteins 0.000 description 9
- 239000012153 distilled water Substances 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 6
- 238000002791 soaking Methods 0.000 description 6
- 239000012564 Q sepharose fast flow resin Substances 0.000 description 5
- 239000012506 Sephacryl® Substances 0.000 description 5
- 238000004440 column chromatography Methods 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 238000010828 elution Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 102000004877 Insulin Human genes 0.000 description 3
- 108090001061 Insulin Proteins 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- 229940125396 insulin Drugs 0.000 description 3
- 150000002772 monosaccharides Chemical class 0.000 description 3
- 230000011506 response to oxidative stress Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000003929 heteronuclear multiple quantum coherence Methods 0.000 description 2
- 238000004192 high performance gel permeation chromatography Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000003914 insulin secretion Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- 208000002249 Diabetes Complications Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 241000736199 Paeonia Species 0.000 description 1
- 241000218201 Ranunculaceae Species 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 238000003570 cell viability assay Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 239000008236 heating water Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- -1 oxygen free radical Chemical class 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0009—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Glucans, e.g. polydextrose, alternan, glycogen; (alpha-1,4)(alpha-1,6)-D-Glucans; (alpha-1,3)(alpha-1,4)-D-Glucans, e.g. isolichenan or nigeran; (alpha-1,4)-D-Glucans; (alpha-1,3)-D-Glucans, e.g. pseudonigeran; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/716—Glucans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Diabetes (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Materials Engineering (AREA)
- Animal Behavior & Ethology (AREA)
- Biochemistry (AREA)
- Emergency Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Sustainable Development (AREA)
- Epidemiology (AREA)
- Endocrinology (AREA)
- Mycology (AREA)
- Hematology (AREA)
- Obesity (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses cortex moutan linear alpha-D-1, 4-glucan and a preparation method and application thereof, and belongs to the technical field of polysaccharide extraction. The molecular weight range of the natural linear alpha-D-1, 4-glucan obtained by the invention is 1.5-3.3kDa, and crude polysaccharide of cortex moutan is obtained by drying and crushing cortex moutan, heating, refluxing and degreasing, extracting with room temperature water, centrifuging, concentrating supernate under reduced pressure, precipitating with ethanol, filtering under reduced pressure, and freeze-drying; and dissolving the crude cortex moutan polysaccharide in water to form a crude cortex moutan polysaccharide solution, adding an organic solvent to remove protein, dialyzing, separating and purifying, concentrating and freeze-drying to obtain the linear alpha-D-1, 4-glucan of cortex moutan. The cortex moutan linear alpha-D-1, 4-glucan has a protective effect on high-sugar-induced oxidative damage of islet beta cells, shows a hypoglycemic effect, and has no toxic or side effect on normal cells.
Description
Technical Field
The invention relates to cortex moutan linear alpha-D-1, 4-glucan and a preparation method and application thereof, belonging to the technical field of polysaccharide extraction.
Background
Diabetes is one of the major chronic diseases seriously harming human health, diabetes and complications thereof become worldwide public health problems seriously harming human health, and high importance is attached to all countries in the world, and most of the diabetics belong to type II diabetes (T2 DM). Oxidative stress is the most fundamental cause of type II diabetes mellitus and complications thereof, and islet beta cell injury caused by oxidative stress due to imbalance of oxidation and antioxidation is an important mechanism for the occurrence and development of diabetes mellitus. Under the conditions of high sugar environment, high fat state, cell factors and the like, the insulin beta cells can be induced to generate oxidative stress reaction, so that the insulin beta cells are damaged, and the insulin secretion is reduced.
At present, except insulin injection, the clinical treatment of the II type diabetes mainly comprises oral hypoglycemic drugs, one part of which is chemically synthesized hypoglycemic drugs, but has the defects of large development difficulty and easy generation of adverse reaction, and in addition, hypoglycemic active ingredients are extracted from natural products. Cortex moutan (Paeonia suffruticosa Andr.) is the dry root bark of Paeonia suffruticosa of Paeonia of Ranunculaceae, and the cortex moutan polysaccharide contained in the cortex moutan is known to have the effect of treating diabetes at present, but the cortex moutan polysaccharide is composed of various monosaccharides such as L-rhamnose, L-arabinose, D-mannose, D-xylose and the like, and how to further extract more targeted polysaccharide components from the cortex moutan polysaccharide has important significance for the development and research of hypoglycemic drugs and functional foods.
Disclosure of Invention
In order to solve the problem that the islet beta cells generate oxidative stress reaction in a high-sugar environment, the invention provides cortex moutan linear alpha-D-1, 4-glucan and a preparation method and application thereof, and the cortex moutan linear alpha-D-1, 4-glucan has a protection effect on the islet beta cells with high-sugar induced oxidative damage, has no toxic or side effect on normal cells, and has an effect of reducing blood sugar.
In order to achieve the purpose, the invention provides the following scheme:
the technical scheme is as follows:
a cortex moutan linear alpha-D-1, 4-glucan has the following structural formula:
[→4)-α-D-Glcp-(1→]n
wherein n is a positive integer, the molecular weight range is 1.5-3.3kDa, and the average molecular weight is 2.4 kDa.
The second technical proposal is that:
a method for preparing cortex moutan linear alpha-D-1, 4-glucan comprises the following steps:
1) drying and crushing cortex moutan, adding 80% ethanol, heating, refluxing, degreasing, extracting, drying at room temperature to obtain cortex moutan degreased powder, then adding water, standing at room temperature, stirring, extracting and centrifuging, collecting supernatant, repeatedly extracting precipitate after centrifuging for 2-3 times, mixing the extracted supernatants, concentrating under reduced pressure, precipitating with ethanol, filtering under reduced pressure, re-dissolving the precipitate in water, and freeze-drying to obtain crude cortex moutan polysaccharide;
2) dissolving the crude cortex moutan polysaccharide obtained in the step 1) in water to form a crude cortex moutan polysaccharide solution, adding an organic solvent for precipitation to remove protein, dialyzing, separating and purifying, concentrating and freeze-drying to obtain the linear cortex moutan alpha-D-1, 4-glucan.
Further, the feed-liquid ratio of the cortex moutan defatted powder to water in the step 1) is 1g:20-60mL, the standing time is 1-6h, and the alcohol precipitation condition is that 2-6 times of volume of absolute ethyl alcohol is added, and the mixture is allowed to stand at 4 ℃ for 12 h.
Further, the volume concentration of the cortex moutan crude polysaccharide solution in the step 2) is 1-5%.
Further, the organic solvent in the step 2) is a mixed solution of dichloromethane and n-butanol, and the volume ratio of the dichloromethane to the n-butanol is 6:1-3: 1.
Further, the volume ratio of the cortex moutan crude polysaccharide solution to the organic solvent in the step 2) is 1: 1-3.
Further, the dialysis in step 2) has a molecular weight cut-off of 1000Da, and is separated by anion exchange resin and purified by gel filtration chromatography.
The third technical scheme is as follows:
the cortex moutan linear alpha-D-1, 4-glucan is applied to the preparation of hypoglycemic drugs or functional foods.
Further, the hypoglycemic drug is used for diabetes caused by the oxidative damage of islet beta cells.
The fourth technical proposal is that:
a pharmaceutical composition comprises the cortex moutan linear alpha-D-1, 4-glucan and one or more pharmaceutically acceptable auxiliary materials.
A health food composition comprises the cortex moutan linear alpha-D-1, 4-glucan and one or more food acceptable auxiliary materials, wherein the cortex moutan linear alpha-D-1, 4-glucan is used as a main component.
The invention discloses the following technical effects:
the invention separates uniform linear alpha-D-1, 4-glucan from cortex moutan, the linear alpha-D-1, 4-glucan can reduce the combined expression of glycosylation products and ROS (reactive oxygen free radical) on islet beta cells under a high-sugar environment, reduce the oxidative stress reaction of the islet beta cells, has a protective effect on the high-sugar induced oxidative damage of the islet beta cells, has no toxic or side effect on normal cells, has the effect of reducing blood sugar, and has important significance for researching novel blood sugar reducing medicines or functional foods.
The cortex moutan polysaccharide prepared by the invention is uniform linear glucan, has a single structure, has an average molecular weight of 2.4kDa, has a small molecular weight, and is convenient to absorb and utilize.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.
FIG. 1 is a high performance gel permeation chromatogram of product A prepared in example 1;
FIG. 2 is a high performance liquid chromatogram of monosaccharide composition of product A prepared in example 1;
FIG. 3 is a NMR chart of product A prepared in example 1;
FIG. 4 is a NMR carbon spectrum of product A prepared in example 1;
FIG. 5 shows the HMQC spectrum of the NMR of product A prepared in example 1;
FIG. 6 is a graph showing the effect of different concentrations of the product prepared in example 1 on the activity of mouse islet beta cell INS-1;
FIG. 7 is a graph showing the effect of different concentrations of the product prepared in example 1 on the protection of high-sugar induced mouse islet beta cell INS-1.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The description and examples are intended to be illustrative only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The percentage of the 80% ethanol is volume fraction unless otherwise specified.
The technical solution of the present invention is further illustrated by the following examples.
Example 1
Drying and crushing tree peony bark, degreasing the obtained powder with 80% ethanol for 24h, drying at room temperature, adding 3000mL of distilled water into 100g of tree peony bark powder, soaking at room temperature for 2h, stirring and extracting for 2h, centrifuging, collecting supernatant, heating the residue, stirring and extracting for 2 times with hot water, centrifuging, mixing the supernatants, concentrating, adding 5 times of volume of absolute ethyl alcohol for alcohol precipitation, precipitating, freeze-drying to obtain tree peony bark hot-water-extracted crude polysaccharide, wherein the polysaccharide yield is 5.6%;
preparing 1% solution from crude polysaccharide of cortex moutan, adding equal volume of dichloromethane/n-butanol mixed solution (the volume ratio of dichloromethane to n-butanol is 4:1), stirring to remove protein, repeatedly removing protein for 2 times, concentrating the polysaccharide solution after protein removal, dialyzing with a dialysis bag with the cut-off molecular weight of 1000Da, concentrating the dialysate, purifying the polysaccharide with Q-Sepharose Fast Flow anion exchange resin, eluting with 0-1mol/L sodium chloride solution, collecting water elution components, concentrating, further purifying with gel column chromatography Sephacryl S-100, and collecting polysaccharide components to obtain product A.
Structural characterization:
the relative molecular weight and purity of product A obtained in example 1 were determined by High Performance Gel Permeation Chromatography (HPGPC) using an Agilent 1260 high performance liquid chromatograph, TSK Gel 3000 as column, 0.1% sodium azide as mobile phase, 1.0mL/min flow rate, 35 ℃. The purity identification result is shown in FIG. 1, and can be seen from FIG. 1, the result is a single symmetrical chromatographic peak, which indicates that the obtained product A is a polysaccharide with uniform structure, and the relative weight average molecular weight is 2.4 kDa.
Component analysis:
PMP pre-column derivatization high performance liquid chromatography for monosaccharide composition analysis shows that the product A mainly consists of glucose, and the result is shown in figure 2.
Detecting correlation between hydrogen signals and carbon signals:
hydrogen nuclear magnetic resonance spectrum (FIG. 3), carbon spectrum (FIG. 4), and1H-13the C HMQC (figure 5) spectrogram can obtain the correlation between hydrogen signals and carbon signals, namely 5.4-100.6(H1/C1), 3.64-72.1(H2/C2), 3.96-74.1(H3/C3), 3.66-77.7(H4/C4), 3.85-71.9(H5/C5), 3.66 and 3.83-61.2 (H6/C6).
As shown by structural characterization, component analysis and detection and analysis of correlation between hydrogen signals and carbon signals, the product A obtained in example 1 is alpha-D-1, 4-glucan with the average molecular weight of 2.4 kDa. The structural formula is [ → 4) -alpha-D-Glcp- (1 →]n。
Example 2
Drying and crushing tree peony bark, degreasing the obtained powder with 80% ethanol for 24h, drying at room temperature, adding 2000mL of distilled water into 100g of tree peony bark powder, soaking at room temperature for 3h, stirring and extracting for 2h, centrifuging, collecting supernatant, heating the residue, stirring and extracting for 2 times with hot water, centrifuging, mixing the supernatants, concentrating, adding 4 times of volume of absolute ethyl alcohol for alcohol precipitation, precipitating, freeze-drying to obtain tree peony bark hot water extraction crude polysaccharide, wherein the polysaccharide yield is 4.8%;
preparing 2% solution from crude polysaccharide of cortex moutan, adding dichloromethane/n-butanol mixed solution with twice volume (the volume ratio of dichloromethane to n-butanol is 6:1), stirring to remove protein, repeatedly removing protein for 2 times, concentrating the polysaccharide solution after protein removal, dialyzing with a dialysis bag with cut-off molecular weight of 1000Da, concentrating the dialysate, purifying polysaccharide with Q-Sepharose Fast Flow anion exchange resin, eluting with 0-1mol/L sodium chloride solution, collecting water elution components, concentrating, further purifying with gel column chromatography Sephacryl S-100, and collecting polysaccharide components to obtain product B.
As shown by structural characterization, component analysis and detection and analysis of correlation between hydrogen signals and carbon signals, the product B obtained in example 1 is alpha-D-1, 4-glucan with the average molecular weight of 2.4 kDa.
Example 3
Drying and crushing tree peony bark, degreasing the obtained powder with 80% ethanol for 24h, drying at room temperature, adding 2500mL of distilled water into 100g of tree peony bark powder, soaking at room temperature for 4h, stirring and extracting for 2h, centrifuging, collecting supernatant, heating the residue, stirring and extracting for 2 times with hot water, centrifuging, mixing the supernatants, concentrating, adding 4 times of volume of absolute ethyl alcohol for alcohol precipitation, precipitating, freeze-drying to obtain tree peony bark hot water extraction crude polysaccharide, wherein the polysaccharide yield is 4.5%;
preparing 5% solution from crude polysaccharide of cortex moutan, adding dichloromethane/n-butanol mixed solution with three volumes (the volume ratio of dichloromethane to n-butanol is 3:1), stirring to remove protein, repeatedly removing protein for 2 times, concentrating the polysaccharide solution after protein removal, dialyzing by using a dialysis bag with the cut-off molecular weight of 1000Da, concentrating the dialysate, purifying the polysaccharide by using Q-Sepharose Fast Flow anion exchange resin, eluting by using 0-1mol/L sodium chloride solution, collecting water elution components, concentrating, further purifying by using gel column chromatography Sephacryl S-100, and collecting polysaccharide components to obtain a product C.
As shown by structural characterization, component analysis and detection and analysis of correlation between hydrogen signals and carbon signals, the product C obtained in example 1 is alpha-D-1, 4-glucan with the average molecular weight of 2.4 kDa.
Example 4
Drying and crushing tree peony bark, degreasing the obtained powder for 24 hours by using 80% ethanol, drying at room temperature, adding 4500mL of distilled water into 100g of tree peony bark powder, soaking at room temperature for 6 hours, stirring and extracting for 2 hours, centrifuging, collecting supernate, heating water, stirring and extracting residues for 2 times, centrifuging, combining supernate, concentrating, adding 2 times volume of absolute ethanol for alcohol precipitation, precipitating, freeze-drying to obtain tree peony bark hot-water-extracted crude polysaccharide, wherein the polysaccharide yield is 4.2%;
preparing 3% solution from crude polysaccharide of cortex moutan, adding equal volume of dichloromethane/n-butanol mixed solution (the volume ratio of dichloromethane to n-butanol is 5:1), stirring to remove protein, repeating protein removal for 2 times, concentrating the polysaccharide solution after protein removal, dialyzing with a dialysis bag with a molecular weight cut-off of 3500Da, concentrating the dialysate, purifying the polysaccharide with Q-Sepharose Fast Flow anion exchange resin, eluting with 0-1mol/L sodium chloride solution, collecting water elution components, concentrating, further purifying with gel column chromatography Sephacryl S-100, and collecting polysaccharide components to obtain product D.
As shown by structural characterization, component analysis and detection and analysis of correlation between hydrogen signals and carbon signals, the product D obtained in example 1 is alpha-D-1, 4-glucan with the average molecular weight of 2.4 kDa.
Example 5
Drying and crushing tree peony bark, degreasing the obtained powder with 80% ethanol for 24h, drying at room temperature, adding 4000mL of distilled water into 100g of tree peony bark powder, soaking at room temperature for 1h, stirring and extracting for 2h, centrifuging, collecting supernatant, heating the residue, stirring and extracting for 2 times with hot water, centrifuging, mixing the supernatants, concentrating, adding 5 times of volume of absolute ethyl alcohol for alcohol precipitation, precipitating, freeze-drying to obtain tree peony bark hot-water-extracted crude polysaccharide, wherein the polysaccharide yield is 4.0%;
preparing 4% solution from crude polysaccharide of cortex moutan, adding dichloromethane/n-butanol mixed solution with twice volume (the volume ratio of dichloromethane to n-butanol is 4:1), stirring to remove protein, repeatedly removing protein for 2 times, concentrating the polysaccharide solution after protein removal, dialyzing with a dialysis bag with cut-off molecular weight of 1000Da, concentrating the dialysate, purifying polysaccharide with Q-Sepharose Fast Flow anion exchange resin, eluting with 0-1mol/L sodium chloride solution, collecting water elution components, concentrating, further purifying with gel column chromatography Sephacryl S-100, and collecting polysaccharide components to obtain product E.
As shown by structural characterization, component analysis and correlation detection analysis of hydrogen signals and carbon signals, the product E obtained in example 1 is alpha-D-1, 4-glucan with the average molecular weight of 2.4 kDa.
Comparative example 1
The only difference from example 1 is that dialysis bag with molecular weight cut-off of 2000Da was used, and the yield of polysaccharide was 1.8%. According to structural characterization, component analysis and correlation detection analysis of hydrogen signals and carbon signals, the final product obtained in comparative example 1 has an average molecular weight of 2.1 kDa.
Comparative example 2
The difference from example 1 is only that crude polysaccharide from cortex moutan was prepared into 6 vol% solution, and the yield of polysaccharide was 2.5%. According to structural characterization, component analysis and correlation detection analysis of hydrogen signals and carbon signals, the final product obtained in comparative example 2 has an average molecular weight of 2.2 kDa.
Comparative example 3
The only difference from example 1 is that 100g of cortex moutan powder was added to 1000mL of distilled water, and the polysaccharide yield was 2.1%. According to structural characterization, component analysis and correlation detection analysis of hydrogen signals and carbon signals, the final product obtained in comparative example 3 has an average molecular weight of 2.3 kDa.
Comparative example 4
The only difference from example 1 is that 95% ethanol precipitation in 7 volumes was performed, resulting in a polysaccharide yield of 4.5%. As shown by structural characterization, component analysis and correlation detection analysis of hydrogen signals and carbon signals, the average molecular weight of the product obtained in comparative example 4 is 3.0 kDa.
Comparative example 5
Drying and crushing tree peony bark, degreasing the obtained powder for 24h by using 80% ethanol, drying at room temperature, adding 3000mL of distilled water into 100g of tree peony bark powder, soaking at room temperature for 2h, stirring and extracting for 2h, centrifuging, removing supernatant, heating the residue, stirring and extracting for 2 times by using hot water, centrifuging, combining the supernatants, concentrating, adding 3 times of 95% ethanol by volume for precipitating, freeze-drying, and obtaining the tree peony bark hot water extracted crude polysaccharide. According to structural characterization, component analysis and detection and analysis of correlation between hydrogen signals and carbon signals, the average molecular weight of the water-extracted crude polysaccharide obtained in the comparative example 5 is 2.2 kDa.
Islet beta cell protection assay:
taking normal mouse islet beta cell INS-1, culturing in a high-glucose DMEM medium containing 10% fetal calf serum at 37 ℃ in an incubator containing 5% carbon dioxide, after the cells were attached to the wall and fully grown, 0.25% trypsin was used for digestion, the cells were inoculated into a 96-well cell culture plate (cell density: 5000 cells/well), the cortex moutan polysaccharide sample (CPA) solution (drug addition group) prepared in example 1 was added after the cells were attached to the wall to make the final concentration of 100, 200, 400, 800 and 1000 μ g/mL, the phosphate buffer solution was added in the negative Control group (Control), no solution was added in the blank group, 3 multiple wells were set, after incubating the cells for 24 and 48h, MTT (thiazole blue) color reagent cells were added for incubation for 4h, the supernatant was extracted, DMSO (dimethyl sulfoxide) was added, and the OD value (cell survival rate) of each well was measured at a wavelength of 570nm using an enzyme reader:
cell survival (%) ═ ODMedicine adding device-ODBlank group]/[ODNegative control group-ODBlank group]
The effect of each group on the cell viability of mouse islet beta cell INS-1 is shown in FIG. 6, and it can be seen from the figure that the cell viability of the product obtained in example 1 is not significantly different in 24h and 48h after the cells are incubated at 100, 200, 400, 800 and 1000. mu.g/mL compared with that of the negative control group, which indicates that the polysaccharide product prepared in the embodiment of the present invention has no toxic and side effects on normal cells at high concentration.
The cortex moutan polysaccharide has the protection effect on high-sugar induced islet beta cell INS-1:
mouse islet beta cell INS-1 is cultured in a high-glucose DMEM culture medium containing 10% fetal calf serum in an incubator containing 5% carbon dioxide at 37 ℃, after the cells grow full of adherent cells, the cells are digested by 0.25% trypsin, the cells are inoculated in a 96-well cell culture plate (the cell density is 5000 cells/well), and after the cells adhere to the wall, 40MM glucose is added for incubation, so that a high-glucose-induced mouse islet beta cell INS-1 model is established.
Then, the cortex moutan polysaccharide (CPA) sample solution (drug addition group) prepared in example 1 was added to make the final concentrations 100, 200, 400, 800 and 1000 μ g/mL, the negative Control group (Control) was phosphate buffer solution, the blank group was not added with solution, 3 duplicate wells were set, after incubating the cells for 48h, MTT was added to incubate the cells for 4h, the supernatant was aspirated, DMSO was added, and the OD value (cell survival rate) of each well was measured at a wavelength of 570nm using a microplate reader:
cell survival (%) ═ ODMedicine adding device-ODBlank group]/[ODNegative control group-ODBlank group]
The results of cell viability assay after the product solution prepared in example 1 was allowed to act on glucose-induced mouse islet beta cells INS-148 h are shown in fig. 7, where # # # is a blank group and # # is an additive group. The cell viability is gradually increased along with the increase of the concentration of the product prepared in example 1, which shows that the alpha-D-1, 4-glucan prepared in example 1 can reduce the cytotoxicity of glucose on mouse islet beta cell INS-1 cells and has a protective effect on oxidatively damaged mouse islet beta cell INS-1 cells, so that insulin secretion is increased to play a role in reducing blood sugar.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (6)
1. The application of the cortex moutan linear alpha-D-1, 4-glucan in preparing hypoglycemic drugs or functional foods, wherein the hypoglycemic drugs are used for treating diabetes caused by the oxidative damage of islet beta cells;
the cortex moutan linear alpha-D-1, 4-glucan has the following structural formula:
[→4)-α-D-Glcp-(1→]n
wherein n is a positive integer, the molecular weight range is 1.5-3.3kDa, and the average molecular weight is 2.4 kDa;
the preparation method of the cortex moutan linear alpha-D-1, 4-glucan comprises the following steps:
1) drying and pulverizing cortex moutan, adding ethanol, reflux-extracting, drying to obtain cortex moutan defatted powder, adding water, standing, stirring, extracting, centrifuging, collecting supernatant, concentrating under reduced pressure, precipitating with ethanol, vacuum filtering, and freeze-drying to obtain cortex moutan crude polysaccharide;
2) dissolving the crude cortex moutan polysaccharide obtained in the step 1) in water to form a crude cortex moutan polysaccharide solution, adding an organic solvent to remove protein, dialyzing, separating and purifying, concentrating and freeze-drying to obtain the linear cortex moutan alpha-D-1, 4-glucan.
2. The application of claim 1, wherein the feed-liquid ratio of the cortex moutan defatted powder to water in step 1) is 1 g/20-60 mL, the standing time is 1-6h, and the alcohol precipitation condition is to add 2-6 times of volume of absolute ethanol and to stand at 4-6 ℃ for 12-15 h.
3. The use of claim 1, wherein the volume concentration of the solution of cortex moutan crude polysaccharide in step 2) is 1-5%.
4. The use according to claim 1, wherein the organic solvent in step 2) is a mixture of dichloromethane and n-butanol, and the volume ratio of dichloromethane to n-butanol is 6:1-3: 1.
5. The use of claim 1, wherein the volume ratio of the solution of crude cortex moutan polysaccharide in step 2) to the organic solvent is 1: 1-3.
6. Use according to claim 1, wherein the dialysis in step 2) has a molecular weight cut-off of 1000Da, separation with an anion exchange resin and purification by gel filtration chromatography.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110883581.8A CN113480677B (en) | 2021-08-03 | 2021-08-03 | Cortex moutan linear alpha-D-1, 4-glucan and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110883581.8A CN113480677B (en) | 2021-08-03 | 2021-08-03 | Cortex moutan linear alpha-D-1, 4-glucan and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113480677A CN113480677A (en) | 2021-10-08 |
| CN113480677B true CN113480677B (en) | 2022-06-07 |
Family
ID=77945272
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110883581.8A Active CN113480677B (en) | 2021-08-03 | 2021-08-03 | Cortex moutan linear alpha-D-1, 4-glucan and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113480677B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114805624B (en) * | 2022-04-29 | 2023-06-20 | 华南理工大学 | Cortex moutan polysaccharide and preparation method and application thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19827978C2 (en) * | 1998-06-24 | 2000-11-30 | Aventis Res & Tech Gmbh & Co | Process for the preparation of water-insoluble alpha-1,4 glucans |
| DE19860373B4 (en) * | 1998-12-28 | 2004-02-19 | Celanese Ventures Gmbh | Oral care products and use of spherical microparticles |
| CA2542249A1 (en) * | 2003-10-24 | 2005-05-06 | Bayer Cropscience Gmbh | Use of linear poly-alpha-1,4-glucans as resistant starch |
| CN101390962B (en) * | 2008-10-15 | 2011-06-22 | 安徽大学 | Preparation method of active components in root-bark tree peony by in-phase leaching and sub-item preparation |
| CN112941127B (en) * | 2021-02-08 | 2023-07-07 | 上海华源制药安徽广生药业有限公司 | Method for extracting paeonol from tree peony bark and combining paeonol with tree peony bark polysaccharide |
-
2021
- 2021-08-03 CN CN202110883581.8A patent/CN113480677B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN113480677A (en) | 2021-10-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108752497B (en) | Preparation of morinda officinalis aqueous extract, oligosaccharide and polysaccharide and application thereof | |
| CN101766678B (en) | Application of total flavonoid in astragalus to preparing medicaments for preventing and controlling diabetes and nephropathy | |
| CN112574326B (en) | Rhizoma gastrodiae macromolecule linear straight-chain glucan and preparation method and application thereof | |
| CN101935359B (en) | Liriope muscari baily polysaccharide and preparation method thereof | |
| CN113480677B (en) | Cortex moutan linear alpha-D-1, 4-glucan and preparation method and application thereof | |
| CN107281258B (en) | Doudouye flavonoid extract and preparation method and application thereof | |
| CN110540603B (en) | Rhizoma anemarrhenae polysaccharide, and preparation method, identification method and application thereof | |
| KR20150055703A (en) | Manufacturing method of small molecule Ginsenoside | |
| CN117281885A (en) | Application of selenium-enriched yam glycoprotein as immunomodulating drug | |
| CN115894735B (en) | Eagle tea acidic polysaccharide, extraction and purification method and anticancer application thereof | |
| CN110218262B (en) | Application of low-sulfated heteroglycan rich in glucuronic acid and derived from brown algae in preparation of medicines for treating type 2 diabetes | |
| KR100316567B1 (en) | Anti-cancer ginseng saponin, method for producing the same, and anticancer composition containing the same as an active ingredient | |
| CN116240256A (en) | Ginseng glycopeptide and preparation process thereof | |
| CN114107418B (en) | Preparation method of ginseng polypeptide | |
| CN114957497A (en) | Gentiana rigescens acidic polysaccharide and preparation method and application thereof | |
| CN112794925B (en) | Amomum villosum polysaccharide and preparation method and application thereof | |
| CN108424469A (en) | Gorgon fruit kernel polysaccharide and separation and extraction method and application thereof | |
| CN113105567A (en) | Paecilomyces cicadae mannan and preparation and application thereof | |
| CN113004299A (en) | Xanthone compound in mangosteen bark for reducing postprandial blood sugar, and extraction method and application thereof | |
| CN105343158A (en) | Fructus sophorae total flavonoid extract with broad-spectrum anti-tumor activity and preparation method and application of fructus sophorae total flavonoid extract | |
| CN111358800A (en) | Application of two phenylethanoid glycoside compounds in preparation of antidiabetic drugs | |
| CN110143989A (en) | A kind of novel Ellagitannins class alpha-glucosidase restrainer and preparation method thereof | |
| CN110882263B (en) | Application of Monostroma nitidum oligosaccharose in preparing anti-breast cancer medicine | |
| CN107802715A (en) | Water chestnut extract with the effect of α Glucosidase inhibitors and its preparation method and application | |
| CN111675772B (en) | Natural component for inhibiting melanin synthesis and application thereof in cosmetics |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |